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    Infection of Biomphalaria glabrata snails with Schistosoma mansoni miracidia

    Sarah K Buddenborg, Matt Berriman, Geetha Sankaranarayanan, Magda E Lotkowska, Catherine McCarthy, Carmen L Dias Soria, Lisa Seymour, Gabriel Rinaldi

    Published: 2023-04-12 DOI: 10.17504/protocols.io.36wgqjkkxvk5/v2
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    Abstract

    To infect Biomphalaria glabrata snails with miracidia hatched from Schistosoma mansoni eggs

    Before start

    Steps

    Liver preparation

    1.

    Collect livers from patent mice into 50ml Falcon tubes containing pre-warmed 37°C 1x DPBS

    2.

    Remove livers from PBS and place in large mortar or laboratory blender

    3.

    Gently homogenise liver tissue

    4.

    Place homogenised liver slurry into a flask (size of flask depends on how many livers you have) and fill with diH2O or aquarium water

    Hatching miracidia

    5.

    Cover flask in aluminum foil and shine light horizontally across the opening of the flask for 1-2 hrs

    6.

    Take a small aliquot of water from the top of the flask using a glass Pasteur pipette and place into a petri dish. Using a stereomicroscope, check for swimming miracidia

    Exposing snails to miracidia for regular life cycle maintenance

    7.

    Under a microscope, collect 15 miracidia with a glass Pasteur pipette per well

    Alternatively (but less preferred) estimate the number of miracidia by counting 12-5µl aliquots in a petri dish. After collecting (shaking the tube in between) and placing the aliquots in the petri dish add 5µl of Lugol to each drop (this kills and stains the miracidia)

    8.

    Place individual 5-8mm snails into 24-well culture plates and cover snails completely with 1x aquarium water

    9.

    Expose snails to miracidia for at least 3 hours (up to overnight)

    10.

    After exposure, remove the snails carefully using featherweight forceps and put them in a new tank with food

    Exposing snails to miracidia for monomiracidium infections

    11.

    Dilute miracidia so that one miracidium can easily be collected in ~3-5µl of water

    12.

    Collected single miracidium using fresh 10µl pipette each collection and place in 24-well plate

    13.

    After a plate is filled, check each well under a microscope to verify there is a single miracidium in each well

    14.

    Place individual 5-8mm snails in the wells containing confirmed single miracidium and cover snails completely with 1x aquarium water

    15.

    Leave the plate overnight (inside incubator or room at 28°C)

    16.

    The following day, transfer the snails to a new tank with food

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